JPH0269155A - Production of soybean curd preservable for a long period at normal temperature - Google Patents
Production of soybean curd preservable for a long period at normal temperatureInfo
- Publication number
- JPH0269155A JPH0269155A JP63219703A JP21970388A JPH0269155A JP H0269155 A JPH0269155 A JP H0269155A JP 63219703 A JP63219703 A JP 63219703A JP 21970388 A JP21970388 A JP 21970388A JP H0269155 A JPH0269155 A JP H0269155A
- Authority
- JP
- Japan
- Prior art keywords
- tofu
- soybean curd
- transglutaminase
- soybean
- retort
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 235000013527 bean curd Nutrition 0.000 title claims abstract description 58
- 238000004519 manufacturing process Methods 0.000 title claims description 8
- 108060008539 Transglutaminase Proteins 0.000 claims abstract description 12
- 102000003601 transglutaminase Human genes 0.000 claims abstract description 12
- 239000000701 coagulant Substances 0.000 claims abstract description 11
- 125000002252 acyl group Chemical group 0.000 claims abstract description 6
- 125000000404 glutamine group Chemical group N[C@@H](CCC(N)=O)C(=O)* 0.000 claims abstract description 4
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 3
- 235000013322 soy milk Nutrition 0.000 claims description 12
- 238000011049 filling Methods 0.000 claims description 3
- 238000006276 transfer reaction Methods 0.000 claims description 3
- 244000068988 Glycine max Species 0.000 abstract description 13
- 235000010469 Glycine max Nutrition 0.000 abstract description 13
- 102000004190 Enzymes Human genes 0.000 abstract description 12
- 108090000790 Enzymes Proteins 0.000 abstract description 12
- 239000000758 substrate Substances 0.000 abstract description 10
- 244000005700 microbiome Species 0.000 abstract description 9
- 235000013336 milk Nutrition 0.000 abstract description 9
- 239000008267 milk Substances 0.000 abstract description 9
- 210000004080 milk Anatomy 0.000 abstract description 9
- 238000006243 chemical reaction Methods 0.000 abstract description 8
- 239000011575 calcium Substances 0.000 abstract description 7
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 abstract description 5
- 241000187747 Streptomyces Species 0.000 abstract description 5
- 229910052791 calcium Inorganic materials 0.000 abstract description 5
- 230000009471 action Effects 0.000 abstract description 2
- 238000012856 packing Methods 0.000 abstract 1
- 230000007704 transition Effects 0.000 abstract 1
- 230000000694 effects Effects 0.000 description 27
- 238000000034 method Methods 0.000 description 19
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 18
- 239000000243 solution Substances 0.000 description 17
- 235000018102 proteins Nutrition 0.000 description 14
- 102000004169 proteins and genes Human genes 0.000 description 14
- 108090000623 proteins and genes Proteins 0.000 description 14
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 12
- 239000007788 liquid Substances 0.000 description 9
- 230000015271 coagulation Effects 0.000 description 8
- 238000005345 coagulation Methods 0.000 description 8
- 238000010438 heat treatment Methods 0.000 description 8
- 239000002253 acid Substances 0.000 description 7
- 238000002360 preparation method Methods 0.000 description 7
- 229910052757 nitrogen Inorganic materials 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 235000002639 sodium chloride Nutrition 0.000 description 6
- 238000003756 stirring Methods 0.000 description 6
- 239000003153 chemical reaction reagent Substances 0.000 description 5
- 238000005259 measurement Methods 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 230000000704 physical effect Effects 0.000 description 5
- AVXURJPOCDRRFD-UHFFFAOYSA-N Hydroxylamine Chemical compound ON AVXURJPOCDRRFD-UHFFFAOYSA-N 0.000 description 4
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 238000002835 absorbance Methods 0.000 description 4
- 239000002518 antifoaming agent Substances 0.000 description 4
- -1 couscougen Chemical compound 0.000 description 4
- 238000001962 electrophoresis Methods 0.000 description 4
- 239000000706 filtrate Substances 0.000 description 4
- 239000007789 gas Substances 0.000 description 4
- YBYRMVIVWMBXKQ-UHFFFAOYSA-N phenylmethanesulfonyl fluoride Chemical compound FS(=O)(=O)CC1=CC=CC=C1 YBYRMVIVWMBXKQ-UHFFFAOYSA-N 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- PHOQVHQSTUBQQK-SQOUGZDYSA-N D-glucono-1,5-lactone Chemical compound OC[C@H]1OC(=O)[C@H](O)[C@@H](O)[C@@H]1O PHOQVHQSTUBQQK-SQOUGZDYSA-N 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 3
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 3
- 101710123874 Protein-glutamine gamma-glutamyltransferase Proteins 0.000 description 3
- 239000007853 buffer solution Substances 0.000 description 3
- 235000014113 dietary fatty acids Nutrition 0.000 description 3
- 239000000839 emulsion Substances 0.000 description 3
- 239000000284 extract Substances 0.000 description 3
- 239000003925 fat Substances 0.000 description 3
- 239000000194 fatty acid Substances 0.000 description 3
- 229930195729 fatty acid Natural products 0.000 description 3
- 150000004665 fatty acids Chemical class 0.000 description 3
- 235000012209 glucono delta-lactone Nutrition 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N hydrochloric acid Substances Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 3
- 239000003112 inhibitor Substances 0.000 description 3
- 230000007774 longterm Effects 0.000 description 3
- 238000000465 moulding Methods 0.000 description 3
- 239000008363 phosphate buffer Substances 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- 238000003860 storage Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- NEAQRZUHTPSBBM-UHFFFAOYSA-N 2-hydroxy-3,3-dimethyl-7-nitro-4h-isoquinolin-1-one Chemical compound C1=C([N+]([O-])=O)C=C2C(=O)N(O)C(C)(C)CC2=C1 NEAQRZUHTPSBBM-UHFFFAOYSA-N 0.000 description 2
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- 241000700199 Cavia porcellus Species 0.000 description 2
- 102000009127 Glutaminase Human genes 0.000 description 2
- 108010073324 Glutaminase Proteins 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 2
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 2
- 235000011130 ammonium sulphate Nutrition 0.000 description 2
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- OSGAYBCDTDRGGQ-UHFFFAOYSA-L calcium sulfate Chemical compound [Ca+2].[O-]S([O-])(=O)=O OSGAYBCDTDRGGQ-UHFFFAOYSA-L 0.000 description 2
- 238000010411 cooking Methods 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 230000001186 cumulative effect Effects 0.000 description 2
- 238000006297 dehydration reaction Methods 0.000 description 2
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 2
- 235000019797 dipotassium phosphate Nutrition 0.000 description 2
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 239000003995 emulsifying agent Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 235000019197 fats Nutrition 0.000 description 2
- 239000012467 final product Substances 0.000 description 2
- 235000013312 flour Nutrition 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 2
- 229910052742 iron Inorganic materials 0.000 description 2
- 238000009630 liquid culture Methods 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 2
- 235000019341 magnesium sulphate Nutrition 0.000 description 2
- 239000003921 oil Substances 0.000 description 2
- 235000019198 oils Nutrition 0.000 description 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 230000001953 sensory effect Effects 0.000 description 2
- 238000002791 soaking Methods 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000000108 ultra-filtration Methods 0.000 description 2
- 239000002023 wood Substances 0.000 description 2
- JIMLDJNLXLMGLX-JTQLQIEISA-N (2s)-5-amino-5-oxo-2-(phenylmethoxycarbonylamino)pentanoic acid Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)OCC1=CC=CC=C1 JIMLDJNLXLMGLX-JTQLQIEISA-N 0.000 description 1
- LDVVTQMJQSCDMK-UHFFFAOYSA-N 1,3-dihydroxypropan-2-yl formate Chemical compound OCC(CO)OC=O LDVVTQMJQSCDMK-UHFFFAOYSA-N 0.000 description 1
- PAWQVTBBRAZDMG-UHFFFAOYSA-N 2-(3-bromo-2-fluorophenyl)acetic acid Chemical compound OC(=O)CC1=CC=CC(Br)=C1F PAWQVTBBRAZDMG-UHFFFAOYSA-N 0.000 description 1
- SOUXAAOTONMPRY-NSHDSACASA-N 2-[[(2s)-5-amino-5-oxo-2-(phenylmethoxycarbonylamino)pentanoyl]amino]acetic acid Chemical compound OC(=O)CNC(=O)[C@H](CCC(=O)N)NC(=O)OCC1=CC=CC=C1 SOUXAAOTONMPRY-NSHDSACASA-N 0.000 description 1
- 241001156739 Actinobacteria <phylum> Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241001131796 Botaurus stellaris Species 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 244000241257 Cucumis melo Species 0.000 description 1
- 235000015510 Cucumis melo subsp melo Nutrition 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 108010024636 Glutathione Proteins 0.000 description 1
- BYYNJRSNDARRBX-YFKPBYRVSA-N Gly-Gln-Gly Chemical compound NCC(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O BYYNJRSNDARRBX-YFKPBYRVSA-N 0.000 description 1
- 101000933604 Homo sapiens Protein BTG2 Proteins 0.000 description 1
- 101000933607 Homo sapiens Protein BTG3 Proteins 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 102100026034 Protein BTG2 Human genes 0.000 description 1
- 102100026035 Protein BTG3 Human genes 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 108010073771 Soybean Proteins Proteins 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 241000209140 Triticum Species 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 241000082085 Verticillium <Phyllachorales> Species 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- FJJCIZWZNKZHII-UHFFFAOYSA-N [4,6-bis(cyanoamino)-1,3,5-triazin-2-yl]cyanamide Chemical compound N#CNC1=NC(NC#N)=NC(NC#N)=N1 FJJCIZWZNKZHII-UHFFFAOYSA-N 0.000 description 1
- 239000006096 absorbing agent Substances 0.000 description 1
- 239000008186 active pharmaceutical agent Substances 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 238000005377 adsorption chromatography Methods 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 229940024606 amino acid Drugs 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 235000019270 ammonium chloride Nutrition 0.000 description 1
- 238000009455 aseptic packaging Methods 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000003610 charcoal Substances 0.000 description 1
- 230000001112 coagulating effect Effects 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 238000012364 cultivation method Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- SWXVUIWOUIDPGS-UHFFFAOYSA-N diacetone alcohol Natural products CC(=O)CC(C)(C)O SWXVUIWOUIDPGS-UHFFFAOYSA-N 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 125000004989 dicarbonyl group Chemical group 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 230000001804 emulsifying effect Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012847 fine chemical Substances 0.000 description 1
- 238000005194 fractionation Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000002523 gelfiltration Methods 0.000 description 1
- 239000000182 glucono-delta-lactone Substances 0.000 description 1
- 229960003681 gluconolactone Drugs 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 229960002989 glutamic acid Drugs 0.000 description 1
- 125000000291 glutamic acid group Chemical group N[C@@H](CCC(O)=O)C(=O)* 0.000 description 1
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 125000003630 glycyl group Chemical group [H]N([H])C([H])([H])C(*)=O 0.000 description 1
- 150000004677 hydrates Chemical class 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- JDNTWHVOXJZDSN-UHFFFAOYSA-N iodoacetic acid Chemical compound OC(=O)CI JDNTWHVOXJZDSN-UHFFFAOYSA-N 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 150000004698 iron complex Chemical class 0.000 description 1
- 238000001155 isoelectric focusing Methods 0.000 description 1
- 230000002045 lasting effect Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 125000003588 lysine group Chemical group [H]N([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])(N([H])[H])C(*)=O 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 150000002739 metals Chemical class 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 239000002736 nonionic surfactant Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- UYWQUFXKFGHYNT-UHFFFAOYSA-N phenylmethyl ester of formic acid Natural products O=COCC1=CC=CC=C1 UYWQUFXKFGHYNT-UHFFFAOYSA-N 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 238000001223 reverse osmosis Methods 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 238000005185 salting out Methods 0.000 description 1
- 238000011218 seed culture Methods 0.000 description 1
- 229920002050 silicone resin Polymers 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- 239000004317 sodium nitrate Substances 0.000 description 1
- 235000010344 sodium nitrate Nutrition 0.000 description 1
- 229940001941 soy protein Drugs 0.000 description 1
- 238000001694 spray drying Methods 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000004575 stone Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
- A23L11/40—Pulse curds
- A23L11/45—Soy bean curds, e.g. tofu
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y203/00—Acyltransferases (2.3)
- C12Y203/02—Aminoacyltransferases (2.3.2)
- C12Y203/02013—Protein-glutamine gamma-glutamyltransferase (2.3.2.13), i.e. transglutaminase or factor XIII
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Botany (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Agronomy & Crop Science (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Beans For Foods Or Fodder (AREA)
Abstract
Description
【発明の詳細な説明】
（ｌ上の利用分野）
本発明は、豆乳液に凝固剤と共に新規トランスグルタミ
ナーゼとを作用させて豆腐を調製し、このようにして得
た豆腐をレトルト処理ヂ等の耐熱容器に充填し、レトル
ト処理してレトルト豆腐を製）Δする方法に関する。Detailed Description of the Invention (Above Field of Application) The present invention involves preparing tofu by allowing soybean milk to react with a coagulant and a novel transglutaminase, and then subjecting the thus obtained tofu to retort processing, etc. It relates to a method of producing retort tofu by filling it into a heat-resistant container and retorting it.
（従来技術、発明が解決しようとする問題点）豆腐の長
期保存法としては、無菌包装がなどが考えられる。しか
し、この方法は特殊な製造環境を心髄とし、かつ、保存
性もチルド保存がベースどなっており日持ちはなお充分
とはいえない。(Prior Art, Problems to be Solved by the Invention) Aseptic packaging may be considered as a long-term preservation method for tofu. However, this method relies on a special manufacturing environment, and its shelf life is still not long enough, as it is based on chilled storage.
このような事情に鑑み、先に、本発明者の一部が発明名
として関与して、ワ腐に冷凍耐性を付与することにより
豆腐の保存性を高める技術を開発したく特公昭５６−３
１９４２ ）。ｌ係る技術により、確かに、保存性は向
上したが、６ケ月以りの長期保存性を付与することは不
可能であった。そこで、本発明は、より簡便に流通に置
くこと、Ｍ朋保存性豆腐の製造法の提供を目的とするも
のである。In view of these circumstances, some of the inventors of the present invention were involved in the invention and wanted to develop a technology to improve the shelf life of tofu by imparting freezing resistance to it.
1942). Although such technology has certainly improved shelf life, it has not been possible to provide long-term shelf life of more than 6 months. Therefore, it is an object of the present invention to provide a method for producing tofu that can be stored more easily and has a longer shelf life.
（問題点を解決づるための手段）
本発明とは、上記問題を解決すべく鋭意研究の結末、Ω
乳液に凝固剤を作用さ１器℃−豆腐を調製するときに、
凝固剤と几にＣａ２＋非依存性でペブヂド鎖内のグルタ
ミン残りのγ−カルボキシアミド阜のアシル転移反応を
触媒する新規［ヘランスグルタミナーげ（以下、Ｂ丁Ｇ
ａ ｓ ｅど１８記することがある。）を信用さＵる
と豆腐に耐レトルト性をｆ４与ザることができ、従って
このようにして調製したΩ腐ｔ、Ｌ耐熱容器に充頃して
レトルト処理に付することによりリトル１〜豆腐とする
ことができることを見出し、本発明を完成した。ここに
、ＢＴＧａｓｅは、新規酵素であって、本発明者の一部
が発明者として関与した発明（特願昭６２−１６５０６
１）に係わるｂので、その酵素特性、製造法等について
は別項に記載する。(Means for solving the problems) The present invention is the result of intensive research to solve the above problems.
When preparing tofu, apply a coagulant to the emulsion.
A novel acyl transfer reaction of the γ-carboxyamide of glutamine in the peptide chain in a Ca2+-independent manner with a coagulant [Herans Glutaminage (hereinafter referred to as B-G)]
There are 18 examples such as a, s, and e. ) can give retort resistance to tofu, and therefore, by filling a heat-resistant container with Ωfu prepared in this way and subjecting it to retort treatment, tofu can be given retort resistance. The present invention was completed by discovering that it can be made into tofu. Here, BTGase is a novel enzyme, an invention in which some of the present inventors were involved as inventors (Patent Application No. 62-16506).
Since b is related to 1), its enzymatic properties, manufacturing method, etc. will be described in a separate section.
本発明によれば、レトルト耐性を右し、常温でＧか月収
上にも及ぶ長期流通可能な豆腐が提供される。According to the present invention, it is possible to provide tofu that has good retort resistance and can be distributed for a long period of time, with a yield exceeding G months at room temperature.
以「、本発明の詳細な説明づる。The following is a detailed description of the present invention.
本発明に従い長期常温保ｑ可能な〜フ腐を製造するに（
よ、まず、豆乳液に凝固剤と１３ＴＱ ａ Ｓ Ｏとを
作用させてレトルトバウヂ簀の耐熱８器に充頑１べさ豆
腐を調製Ｊる。この調製は、凝固剤にｒ３ Ｔ Ｇ ａ
ｓ ｅを併用Ｊること、及びこの（ＪＩ用に伴−う呂
十の７Ｉ、Ｉ＋杓の他は、全て公知の豆腐の調整法に従
って」；い。To produce fufu that can be kept at room temperature for a long time according to the present invention (
First, prepare soybean tofu by treating the soybean milk with a coagulant and 13TQ a SO and pouring it into a heat-resistant retort container. This preparation uses r3 T Ga in the coagulant.
s and e in combination, and this (for JI, 7 I, I + ladle), all other steps are in accordance with known tofu preparation methods.
従って、豆乳液どしては、従来豆腐の調製に用いられて
いる次のような豆乳液を例として挙げることができる。Therefore, examples of soybean milk liquids include the following soymilk liquids conventionally used in the preparation of tofu.
その１は、丸大豆から得られる豆乳液であって、これは
、丸大豆を水浸漬しく望ましくは水温５−３０℃で８〜
２４時間）、磨砕し、ｍホしく浸漬時の吸水吊し含めて
原料大豆の７〜８倍が望ましい）、消泡剤をＩｔＣＩえ
（食用のもの各種、例えば、脂肪酸モノグリレリド、シ
リコン樹脂製）、加熱・蒸煮しく豆乳中にうまく蛋白質
、脂肪分を溶は出さけると共に殺菌とインヒビターの失
活とを行なうために各種機器で、例えば５分かけて１０
０℃まで上げ、そのままの温度で３〜５分保つ）、絞っ
てＡカラを分離除去することによって（ｑることができ
る。その２は、全脂豆乳粉末から得られる豆乳液であっ
て、これは、豆乳粉末に加水しく７〜１５倍吊、好まし
くは９〜１１倍吊）、加熱・撹Ｉ′￥しく２〜１０分か
けて １００℃近辺とし、そのまま２〜１０分保つ）、
放冷することによって得ることができる。その３は、分
離大豆タンパクから得られる豆乳液であって、これは、
タンパク合印例えば５０％以上の脱脂タンパク粉末に水
、例えば植物油等の油脂、必要に応じてデンプン、及び
各種乳化剤を加えて加熱乳化することによって（りるこ
とができる。The first is a soybean milk liquid obtained from whole soybeans, which is prepared by soaking whole soybeans in water, preferably at a water temperature of 5-30°C.
24 hours), grind it (preferably 7 to 8 times the amount of raw soybeans, including water absorption during soaking), and add an antifoaming agent (various edible ones, such as fatty acid monoglyrelide, silicone resin), ), soybean milk is heated and steamed to dissolve the protein and fat in the soymilk, as well as to sterilize and deactivate the inhibitors.
The second method is soy milk liquid obtained from full-fat soy milk powder. This is done by adding water to soy milk powder (7 to 15 times higher, preferably 9 to 11 times higher), heating and stirring (heating and stirring for 2 to 10 minutes to bring it to around 100°C, and keeping it that way for 2 to 10 minutes).
It can be obtained by cooling it. Part 3 is soy milk liquid obtained from isolated soy protein, which is
The protein signature can be removed, for example, by adding water, oil such as vegetable oil, starch, and various emulsifiers as necessary to 50% or more defatted protein powder and emulsifying the mixture by heating.
さて、従来の豆腐の調製では、このような豆乳液に、ｌ
ｉｌＲＭカルシウムの各種水和物、塩化カルシウム、塩
化マグネシウム、天然ニガリ、グルコノデルタラフ１〜
ン（Ｇ Ｄ Ｌ ）等の凝固剤を濃度０．１〜・５％ど
なるように加えｔ’ ｌ’！拌、放置し、Ω乳液を凝固
さけ、適宜脱水して所望の硬さの豆腐を調製するが、本
発明でルよ、この凝固処１！１１の際に、凝固剤とども
にＢＴＧａＳｅを作用させるのである。Now, in conventional tofu preparation, 1 l is added to such soy milk liquid.
various hydrates of ilRM calcium, calcium chloride, magnesium chloride, natural bittern, glucono delta rough 1~
Add a coagulant such as GDL to a concentration of 0.1 to 5%, then t'l'! Tofu with the desired hardness is prepared by stirring and standing, avoiding coagulation of the Ω emulsion, and dehydration as appropriate.In the present invention, during the coagulation step 1!11, BTGaSe is added to the coagulant. Let it happen.
ＢＴＧａｓｃ併用の目的は、そのタンパク架橋高分子化
能を活用し、耐しｉ−ル１〜ｆｌをイ１する豆腐を調製
することにあるが、そのために番よ次のＪ：うな凝固処
理条件を採用すればよい、：
） 〜ｊ乳乳液タンパク製濃度：３〜１０％、好ましく
は４〜７％、
） 凝固剤ニ一般に豆腐調製に用いられるしの全て、好
ましくはＧＤＬ、
ｉｉｉ ） ［３Ｔ Ｇ ａ ｓ ｅ ｉ１１度：
０．１〜１００ ｕ／！７タンバク、好ましくは１〜
１０ＬＪ／ｒＪタンパク、＋ｖ） 凝固温度： ８０
℃以１Ｚ１好ましくは５０〜７０″′Ｃ℃＝＝
■）凝固反応の時間：１０分〜１夜、好ましくは３０〜
１２０分。The purpose of using BTGasc in combination is to utilize its ability to polymerize protein cross-linking to prepare tofu with a resistance of 1 to 1. To achieve this, the following coagulation treatment conditions are required. It is sufficient to adopt: ) ~ j Milk emulsion protein concentration: 3 to 10%, preferably 4 to 7%, ) Coagulant, all of the ingredients commonly used in tofu preparation, preferably GDL, iii) [3T G a s e i 11 degrees:
0.1~100 u/! 7 tanbak, preferably 1~
10LJ/rJ protein, +v) Coagulation temperature: 80
℃ or less 1Z1 Preferably 50-70''C℃== ■) Time of coagulation reaction: 10 minutes to 1 night, preferably 30 to
120 minutes.
な（１３、凝固処理時に本発明の効果を阻害しない範囲
ひ従東豆腐の調製に使用されている乳化剤等の各種添加
物を加えてもよいことは勿論である。(13) It goes without saying that various additives such as emulsifiers used in the preparation of Tofu tofu may be added to the extent that they do not impede the effects of the present invention during the coagulation process.
まｔご、凝固処ｙＪ！後、）疑固液ｔ、ｔ ｉｒ’過脱
水して所望の硬さの豆腐とし、或いは放置、冷７．ＩＩ
又はインキュベージ」ンを行なって豆腐を調製する。い
ずれにしろ、凝固処理によって１９られる凝固液の処理
も、従来のそれでよい。尚、インＶ」、ベーションは、
５〜６０℃で１・〜２４時間行なうとよく、この操作を
行うと、更に保形性と保水性が向上づる。Matgo, coagulation treatment yJ! 7.) After) pseudo-solid liquid t, tir' super-dehydration to obtain tofu of desired hardness, or leave to cool. II
Alternatively, tofu is prepared by performing incubation. In any case, the treatment of the coagulating liquid 19 during the coagulation process may be conventional. In addition, InV", vation is,
It is preferable to carry out the process at 5 to 60°C for 1 to 24 hours, and by carrying out this operation, the shape retention and water retention properties are further improved.
次に、このＪ：うにして調製した豆腐をレトルト処理ヂ
等の耐熱容器に充填し、レトルト処理して最終製品とす
る１６ｏこのようにして調製した豆腐（Ｊ、レトルト耐
性をｔｒする。レトルト処理における加熱条件（よ、８
０〜１３０℃好ましくは１００〜ｌ ２ ！ｉ ’ｌ。Next, the tofu prepared in this manner is filled into a heat-resistant container such as a retort treatment container and subjected to retort treatment to produce the final product. Heating conditions in treatment (Y, 8
0-130°C, preferably 100-12! i'l.
衿
ζ・５〜１２０のりＹましくけ１ｏ・−３０どザればよ
い。尚、他には最終製品化には特別の制限がない。但し
、実際のし１−ル１−処理は「０埴でコン１−ロールさ
れ、いて所定数の微生物を死滅させるのに要覆る最小加
熱１１．１間（分１秒）で・あ−、）”Ｃ、通常２５０
下（１２１，１℃）での最小加熱致死時開（Ｆｏ’）を
指１ 、この値は食品の加熱殺菌効果を表示覆る指標ど
して用いられている。Collar ζ - 5 to 120 glue Y makake 1o -30 is enough. Note that there are no other special restrictions on the final product. However, in actual treatment, the minimum heating time required to kill a predetermined number of microorganisms is 11.1 minutes (1 minute and 1 second). )”C, usually 250
This value is used as an indicator to indicate the effectiveness of heat sterilization of foods.
このようなしトル１〜処理条件を採用する理由は、帛温
で６ケ月以上の保存を品質およびｔｆｔｉ生上可化上可
能ためである。The reason for adopting such treatment conditions is that storage at room temperature for 6 months or more is possible in terms of quality and viability of TFTI.
従ってこのようにして製品されるしｌ〜ル１−豆腐は、
常温で６か月収上にも及ぶ長期流通が可能である。Therefore, the tofu produced in this way is:
It can be distributed for a long period of time, lasting up to 6 months at room temperature.
（本発明の新規トランスグルタミノーピＲＴ Ｇ ａ
ｓ ｅ ）
（１）１−ランスグルタミナーゼとその由来１〜ランス
グルタミナーピ（以下、ＴＧａｓｅと略称ηることがあ
る。）は、ベブ天ド鎖内にあるグルタミン残基のγ−カ
ルボキシアミド基のアシル転移反応を触９Ｘする酵素で
ある。このＴＧａｓｅ（よ、アシル受容体としてタンパ
ク質中のリジン残ＩＡのε−アミノ基が作用すると、分
子内及び分子間にε−（γ−Ｇｌｕ）−［ｙｓ架橋結合
が形成される。また水がアシル受容体として機能すると
きは、グルタミン残基が脱アミド化されグルタミン酸残
Ｖになる反応を進行させる酵素である。(Novel transglutaminopi RT Ga of the present invention
s e ) (1) 1-Lance glutaminase and its origin 1-Lance glutaminase (hereinafter sometimes abbreviated as TGase) is a γ-carboxyamide group of a glutamine residue in the bebendo chain. It is an enzyme that catalyzes the acyl transfer reaction of When the ε-amino group of the lysine residue IA in the protein acts as an acyl acceptor, ε-(γ-Glu)-[ys crosslinks are formed within and between molecules. When it functions as an acyl acceptor, it is an enzyme that proceeds with the reaction in which glutamine residues are deamidated to become glutamic acid residues V.
ＴＧＥＩ Ｓ ｅのこのような性質により、ＴＧａＳＣ
を用いてタンパク含有溶液又はスラリーをゲル化さＵる
ことができる。Due to these properties of TGEIS, TGaSC
can be used to gel a protein-containing solution or slurry.
Ｔ Ｇ ａ Ｓ ｅは、これまでモルモット肝由来のも
の（以１ζ、ＭＴＧａｓｅと略記することがある。）な
どの動物由来のものが知られているが、動物由来のもの
は、安価にまた大量に入手づるのが困難であり、タンパ
クｖＴをゲル化するときは酵木溌度ａ３よび阜？、ｔ
ｆａ度を共に高くする必要があり、またＣａ２４依存性
であるので用途が制限される。TGaSe has been known to be derived from animals such as guinea pig liver (1ζ, sometimes abbreviated as MTGase), but animal-derived products can be produced at low cost and in large quantities. It is difficult to obtain protein vT, and when gelling protein vT, it is necessary to use fermented wood vigor A3 and fu? ,t
It is necessary to increase the fa degree in both cases, and the use is limited because it is Ca24 dependent.
本発明′Ｃ使用する新規［・ランスグルタミナーゼ（Ｂ
ＴＧａｓｅ）は、微生物、例えば、ストレプトベルチシ
リウムｊｍｌの菌により産生きれるものであるが、微生
物由来のＴＧａｓｅについての報告は現時点ではない。The present invention uses a novel transglutaminase (B
TGase) can be produced by microorganisms, such as Streptoverticillium jml, but there are currently no reports on TGase derived from microorganisms.
本発明で使用する微生物由来の［３ＴＧａｓｃは安価に
供給され、かつｖｉ′！Ａ、６容易であるので実用性が
人である。また、ＲＴＧａｓｅを用いることにより、カ
ルシウム非存在下又カルシウム（ｊ在下のいずれでも酵
素（ＢＴＧａｓｅ）濃度及び基質温度が非常に低いとこ
ろ（）で品質の優れたゲル化物を製造できるという利点
がある。The microorganism-derived [3TGasc used in the present invention is inexpensively supplied, and vi'! A.6 It's easy, so it's practical. Further, by using RTGase, there is an advantage that a gelled product of excellent quality can be produced at a place where the enzyme (BTGase) concentration and substrate temperature are very low () either in the absence of calcium or in the presence of calcium (j).
■［３Ｔ Ｇ ａ ｓ ｅのＭ ｉｌ
Ｇ Ｔ Ｇ ａ ｓ ｃを産生する微生物は、例えば、
ス１ヘレブトベルヂシリｆンム・グリピオカルネウム（
Ｓ ｔｒｃｐｔｏｖｃｒｔ＋ｃ１１１＋ｕｍ ｇｒ＋ｓ
ｅｏｃａｒｎｅｕｍ） Ｉ Ｆ ０１２７７６、ス１
−レブ１〜ベルチシリウム・シブモネウム・４ノブ・エ
スピー・シｊ七−ネウム（Ｓ ｔｒｃｐｔｏｖｅｒｔｉ
ｃｉｌｌｉｕｍ ｃｉｎｎａｍｏｎｅｕｍ ｓｕｂ Ｓ
Ｄ 。■ [Microorganisms that produce 3T Gas are, for example,
S1 Helebutoverdicillinum Gripiocarneum (
S trcptovcrt+c111+um gr+s
eocarneum) I F 012776, S1
-Reb 1 ~ Verticillium sibmoneum 4 knob sp.
cillium cinnamoneum sub S
D.
ｃｉｎｎａｍｏｎｅｕｍ） ｌ ＦＱ １２８！ｉ２、
スｌ−レブトベルチシリウム・モバラー丁二ンス（３ｔ
ｒｅｐｔＯＶ（！ｒｔｉｃｉｌｌｉｔ１ｍｍｏｂａｒａ
ｅｎｓｅ） ｌ ＩＴ Ｑ １３８１９笠があげられ
る。cinnamoneum) l FQ 128! i2,
Sl-Levtoverticillium Mobara 2nds (3t
reptOV(!rticillit1mmobara
ense) l IT Q 13819 Kasa is mentioned.
これら微生物を培養し、トランスグルタミナーゼを取Ｖ
１するための培酋法及び精製法等は次の通りである。Cultivate these microorganisms and extract transglutaminase.
The cultivation method, purification method, etc. for this purpose are as follows.
１８ｇ形態としては、液体培養、固体培養いずれも可能
であるが、工業的には深部通気撹拌１８養を１ｊうのが
有利である。又、使用する培養源とじて（よ、一般に微
生物培養に用いられる炭木源、窒素１１１ｊ：ｊ、無８
１１塩及びイの他の微吊栄航源の他、ス１−レブ１〜ベ
ルヂシリウム属に属する微生物の利用出来る・′Ｘ：首
椋て・あれば仝（使用出来る。」８地の炭木源としては
、ブドウ糖、ショ糖、クスクーゲン、グリセリン、デキ
ストリン、澱粉四の伯、脂肪酸、油脂、イ］（幾酸など
が単独で又は組合Ｕて用いられるＵ窒素源としては、照
磯窒素源、石機窒々；踪のいずれも使用可能であり、無
）幾窒素源どしては硝酸アンモニウム、硫酸アンモニウ
ム、ｉｌ累、ｌＩｌ’ｌ酸ソーダ、塩化アンモニウム等
が挙げられる。叉、イ１改窒素源としてｔよ大豆、米、
トウゼロコシ、小麦などの粉、糠、脱脂粕をはじめ］−
ンスｊイーブリカー、ペプトン、肉エキス、カビイン、
アミノ酸、酵１ｕエキス等が挙げられる。無８３１ 堪
及び微ψ栄養素としては、リン酸、マグネシウム、カリ
ウム、鉄、カルシウム、亜鉛等のｊ８類の他ビタミン、
非イオン界面活性剤、消泡剤等の菌の生育や１３丁Ｇ
ａ ｓ（３の産生を促）Ｗするしのであれば必バに応じ
て使用出来る。As for the 18g form, both liquid culture and solid culture are possible, but from an industrial perspective, it is advantageous to use 18g of deep aeration stirring. In addition, as for the culture source used (charcoal wood source generally used for microbial culture, nitrogen 111j:j, no 8
11 In addition to salt and other microorganisms belonging to the genus Verdicillium, microorganisms belonging to the genus Verdicillium can be used. Examples of nitrogen sources include glucose, sucrose, couscougen, glycerin, dextrin, starch, fatty acids, fats and oils, and nitrogen sources used alone or in combination such as Teruiso nitrogen source, Any of the following nitrogen sources can be used, and examples of nitrogen sources include ammonium nitrate, ammonium sulfate, sodium nitrate, sodium chloride, and ammonium chloride. As for soybeans, rice,
Including corn flour, wheat flour, bran, and defatted lees]
nsj e-liquor, peptone, meat extract, kabiin,
Examples include amino acids, yeast 1u extract, and the like. No. 831 Nutrients include phosphoric acid, magnesium, potassium, iron, calcium, zinc, etc., as well as vitamins,
Non-ionic surfactants, anti-foaming agents, etc. for bacterial growth and 13-G
As (promotes the production of 3) W, it can be used as needed.
培養（よ好気的条件て゛、培Ｎ ｆｒＡ度ｔま菌が発育
しＢ Ｔ Ｇ ａ ｓ ｅが産生する範囲であれば良く
、好ましく ＋、１２ ！ｌ ”□ ３ ｂ℃て゛ある
。培養時間は、条件により異なるが、ＢＴＧａｓｅが最
す産生される時間まで培丑すれば良く、通常２〜４日程
度である。Cultivation (more aerobic conditions, preferably within a range where bacteria can grow and BTGase can be produced), preferably 12°C.Culture time Although it varies depending on the conditions, it is sufficient to culture until the time when BTGase is produced the most, which is usually about 2 to 4 days.
Ｂ −Ｔ Ｇ ａ ｓ ｅは液体培養ではｒＱ？！を液
中に溶解されてｄ５す、培養終了後培益液より固形分を
除いた培養ろ液より採取される。Is B-T Gas rQ in liquid culture? ! After the culture is completed, the solids are removed from the culture filtrate and collected from the culture filtrate.
培養ろ液よりＢ Ｔ Ｇ ａ Ｓ ｅを精製するには、
通常酵素精製に用いられるあらゆる方法が使用出来る。To purify BTG a Se from the culture filtrate,
Any method commonly used for enzyme purification can be used.
例えば、エタノール、アセトン、イソプロピルアルコー
ル等の有機溶媒による処理、硫安、な塩等により塩析、
透析限外ろ適法、イオン交換クロア１〜グラフイー、吸
着クロマトグラフィー、ゲルろ過、吸石剤、等電点分画
等の方法が使用出来る。又、これらの方法を適当に組合
せる事によりＢ Ｔ Ｇ ａ ｓ ｃの精製度が上る場
合は適宜組合して行う事が出来る。これらの方法によっ
て得られるＭ本は、安定化剤として各種の塩ｎ１、糖類
、蛋白質、脂質、界面活性剤等を加え或いは加えること
なく、限外ろ過濃縮、逆浸透濃縮、減圧乾燥、凍結乾燥
、噴霧乾燥の方法により液状又は固形のＢＴＧａｓｅを
得ることが出来る。For example, treatment with organic solvents such as ethanol, acetone, and isopropyl alcohol, salting out with ammonium sulfate, salts, etc.
Methods such as dialysis ultrafiltration, ion exchange chromatography, adsorption chromatography, gel filtration, stone absorbing agents, and isoelectric point fractionation can be used. Further, if the degree of purification of BTG asc can be increased by appropriately combining these methods, the methods can be combined as appropriate. M bottles obtained by these methods can be subjected to ultrafiltration concentration, reverse osmosis concentration, vacuum drying, freeze drying, with or without the addition of various salts, sugars, proteins, lipids, surfactants, etc. as stabilizers. Liquid or solid BTGase can be obtained by spray drying.
［３ＴＧａｓｅの活性測定はベンジルオＡ−ジカルボニ
ル−［−グルタミニルグリシンとヒト［１キシルアミン
を基質としてＣａ２４非存在Ｆで反応を行い、生成した
ヒドロキサム酸をトリクロロ酢酸存在°ｌ・で鉄錯体を
形成さｔ！５２！１ｎＩｌｌの吸収を測定し、ヒドロキ
サム酸の聞を検ｍ線より求め活性を粋出する。[3TGase activity measurement is performed by reacting benzyl-A-dicarbonyl-[-glutaminylglycine with human [1-xylamine] as a substrate in F in the absence of Ca24, and forming an iron complex with the generated hydroxamic acid in the presence of trichloroacetic acid. Sat! The absorption of 52!1 nIll is measured, and the activity of hydroxamic acid is determined from the m-line.
Ｂ Ｔ Ｇ ａ ｓ ｃ粘性は、特に記載しないかぎり
以下に記載する方法により測定した。BTGasc viscosity was measured by the method described below unless otherwise specified.
〈活性測定法〉
試薬へ （１，２Ｍ Ｉ−リス塩酸緩衝液（ｐｔ−１
６，０）０１Ｍヒドロキシルアミン
０、ＱＩ Ｍ還元型グルタチオン
０．０３ Ｍベンジルオーキシカルボニルし一グルタミ
ニルグリシン
試薬Ｂ ３Ｎ−塩酸
１２％−１−リクロロ酢酸
５％Ｆ ｅ（４ ６Ｈ２ ０ （ ０．ＩＮ
−トＩＣでに溶解）
上記溶液の１：１：１の混合液を試薬Ｂとする。<Activity measurement method> To the reagent (1,2M I-Lis-HCl buffer (pt-1
6,0) 01M hydroxylamine 0, QI M reduced glutathione 0.03 M benzyloxycarbonyl monoglutaminylglycine Reagent B 3N-hydrochloric acid 12%-1-lichloroacetic acid 5% Fe(4 6H20 (0. IN
(Dissolved in IC) A 1:1:1 mixture of the above solutions was used as Reagent B.
酵素液の０． ０５−に試薬へ〇．５−を加えて混合し
３７°Ｃで１０分間反応（ｐ、試薬Ｂを加えて反応停止
とＦ ｃ Ｉｆ体の形成を行った後５２５ｎＩｌｌの吸
光度を測定覆る。対照としてあらかじめ熱失活さＵたＶ
Ｉ素液を用いて同様に反応さしたものの吸光瓜を測定し
、酵素液との吸光度差を求める。別に！！系液のかわり
にＬ−グルタミン酸γーモノヒトｒｌ：＄号ム酸を用い
て検１ｉ１ Ｐｉｌを作成し、前記吸光度差より生成さ
れたビド［１１トナム酸の早を求め、１分間に１μしル
のヒト［」キサム酸を生成ツる酵素粘付を１単位とした
。0 of the enzyme solution. 05- to reagent 〇. Add 5-, mix, and react at 37°C for 10 minutes (p. Add reagent B to stop the reaction and form F c If body, then measure the absorbance of 525 nIll. As a control, heat-inactivated U Ta V
A similar reaction was carried out using the base solution I, and the absorbance of the melon was measured to determine the difference in absorbance from the enzyme solution. Especially! ! A test 1i1 Pil was prepared using L-glutamic acid γ-monohuman rl:$1000 acid instead of the system solution, and the speed of the generated bido[11tonamic acid was determined from the difference in absorbance, and the rate of 1 μl per minute was determined. The enzyme stickiness that produces human [xamic acid] was defined as one unit.
（３） Ｂ Ｔ Ｇ ａ ｓ ｃ （Ｊ） Ｗ 累Ｒ
性−Ｌのようにして得られる精製１３丁Ｇ ａ Ｓ Ｑ
、即らストレブトベヂシリウム・モバランスＩＦ０１３
８１９の１〜ランスグルタミナーピＩＴＧ−１と命名）
、ストレプトベルチシリウム・グリレオカルネラム（
Ｆ ０ １２７７Ｇの１〜ランスグルタミナーピ（ＢＴ
Ｇ２と命名）、ストレプトベルチシリウム八・シナモネ
ーンム・すプ・エスピー・シプモネウムＩ ｔ” ０
１２８５２のトランスグルタミナーゼ（ＢＴＧ−３と命
名）についてのＭ’ｌＡ化学的性質は次の通り。(3) B T G a sc (J) W Cumulative R
Purified 13 pieces obtained as in G-L G a S Q
, namely Strebtovedicillium mobalans IF013
819-1 ~ Lance Glutaminapi ITG-1)
, Streptoverticillium glileocarnelum (
F 0 1277G 1 ~ Lance Glutamine Napi (BT
G2), Streptoverticillium VIII. Cinnamoneum sp. Cypmoneum I t" 0
The M'lA chemistry for the 12852 transglutaminase (designated BTG-3) is as follows.
ａ）〒適ｐ）１：
基質としてベンジルオキシカルボニル−ルタミニルグリ
シンとヒドロキシルアミンを使用した場合、３７℃、１
０分反応で、１３ＴＧ−１の至適ｔ）Ｈは６〜７にあり
、１３ Ｔ Ｇ − ２の至適ｐＨは６〜７伺近にあり
、Ｂ Ｔ Ｇ − ３の至適ｐＨは６〜フイ」近にある
。a) 〒Applicable p) 1: When benzyloxycarbonyl-lutaminylglycine and hydroxylamine are used as substrates, 37°C, 1
In a 0 minute reaction, the optimal t)H of 13TG-1 is around 6-7, the optimal pH of 13TG-2 is around 6-7, and the optimal pH of BTG-3 is around 6. ~Fui” is nearby.
ｂ）至適温度：
基質としてベンジルオキシカルボニル−し−グルタミニ
ルグリシンとヒドロキシルアミンを使用した場合、ｐｔ
−＋６、１０分反応で、ＢＴＧ−１の至適温度は５５℃
付近であり、ＢＴＧ−２の至適温度は４５℃付近であり
、ＢＴＧ−３の至適温度は４５℃イ」近にある。b) Optimum temperature: When benzyloxycarbonyl-glutaminylglycine and hydroxylamine are used as substrates, pt
-+6, 10 minutes reaction, optimal temperature of BTG-1 is 55℃
The optimal temperature for BTG-2 is around 45°C, and the optimal temperature for BTG-3 is around 45°C.
ｃ） ｐＨ安定性：
３７℃、１０分間処理で、ＢＴＧ−１はｐ１］５・〜９
Ｃ安定であり、Ｂ Ｔ’ Ｇ − ２はｐｔ−４ ５＝
９で安定であり、［３ ｒＧ−３はＩ）Ｈ ６〜っで安
定である。c) pH stability: After treatment at 37°C for 10 minutes, BTG-1 had p1]5.~9
C stable, B T' G − 2 is pt−4 5=
9, and [3 rG-3 is stable at I)H6~.
ｄ）温度安定性：
ｐ１１７で１０分間処理では、ＢＴＧ− １は４０℃で
は８８％活性が残存し、５０℃では７４％活性が残（？
シ、ＢＴＧ−２は４０℃では８６％活性が残存し、５０
℃では５６％活性が残存し、ＢＴＧ−３ｉよ４０℃で８
０％活性が残存し、５０℃では５３％活性が残存する。d) Temperature stability: After treatment with p117 for 10 minutes, BTG-1 had 88% activity remaining at 40°C and 74% activity remaining at 50°C (?
BTG-2 has 86% activity remaining at 40°C, and 50% activity remains at 40°C.
56% activity remained at 40°C, and 8% activity remained at 40°C compared to BTG-3i.
0% activity remains and at 50°C 53% activity remains.
Ｏ）基質特異性：
各Ｂ ’ｒ Ｇ ａ ｓ ｅを用い、各種合成基質とヒ
ドロキシルアミンとの反応を調べた。いずれのＢＴＧａ
ｓｅも合成ｌｉがベンジル第１−ジカルボニルアスパラ
ギニルグリシン、ベンジルオキシカルボニルグルタミン
、グリシルグルタミニルグリシンの場合反応しない。し
かし合成基質がベンジルオキシ力ルポニルグルタミニル
グリシンの場合の反応性は最も高い。この時の各種合成
基質ｍ度は５ｍＭとし！、：。結末ｔよ表−１に示され
る。O) Substrate specificity: Reactions between various synthetic substrates and hydroxylamine were investigated using each B'r Gas. Which BTGa
Se also does not react when synthetic li is benzyl 1-dicarbonylasparaginylglycine, benzyloxycarbonylglutamine, or glycylglutaminylglycine. However, the reactivity is highest when the synthetic substrate is benzyloxyluponylglutaminylglycine. The concentration of various synthetic substrates at this time is 5mM! , :. The outcome t is shown in Table-1.
４Ｔお、表−１中のＣＢＺはベンジル第１ジカルボニル
ｊＪの略であり、０１ｎはグルタミル措の略であり、Ｇ
ＩＶはグリシル基の略であり、△ｓｐはアスパラ１゛ニ
ル括の略である。4T, CBZ in Table 1 is an abbreviation for benzyl primary dicarbonyl jJ, 01n is an abbreviation for glutamyl dicarbonyl, and G
IV is an abbreviation for glycyl group, and Δsp is an abbreviation for aspara-1'nyl group.
表−１
［）金属イオンの影響：
活性測定系に １ｍＭ１ｌ！度になるように各種金属イ
Ａンベハ１１え″（影響を調べたく結末（、（表−２に
示されル）。イヂレノｔ３１−Ｇａ ｓ ｃ ｂＣＬＪ
２”７ｎ２＋Ｍより活性が阻害される。Table-1 [) Effect of metal ions: 1mM1l for activity measurement system! I wanted to investigate the effects of various metals on the substrate 11'' (as shown in Table 2).
The activity is inhibited by 2”7n2+M.
表−２
す）阻害剤の影′ｆＪ：
各阻害剤を１ ｍＭになるように加え、２５℃、３０分
放置後、活性を測定した（結末は人−３に示される）。Table 2) Shadow of inhibitor'fJ: Each inhibitor was added to a concentration of 1 mM, and after standing at 25°C for 30 minutes, the activity was measured (results are shown in person-3).
いずれの（３７Ｑａｓｅもバラク００マー１−Ｉり一安
忠香酸（ＰＣＭｒ３と略する）、Ｎ−ニブルマレイミド
（ＮＥＭと略する）、モノヨード酢酸により活性が阻害
される。The activity of 37Qase is inhibited by 1-I monobenzoic acid (abbreviated as PCMr3), N-nibrumaleimide (abbreviated as NEM), and monoiodoacetic acid.
表−３
表−３中ＰＭＳＦはフェニルメチルスルホニルフルオラ
イドの略である。Table 3 In Table 3, PMSF is an abbreviation for phenylmethylsulfonyl fluoride.
ｈ）等電点：
アン小ライン等電点電気泳動にＪ、り求めたところ、１
１ Ｔ Ｇ−１０秀’；’＋１ ｉ：、ミｌ）Ｉは９イ
・Ｉ近で・あり、ＢＴＧ−２の等電点ｐｉは９フイ・Ｊ
近であり、ＢＴＧ３の等″心慮ｏｆは９８句近である。h) Isoelectric point: J was determined by Anne small line isoelectric focusing electrophoresis, 1
1 T G-10 Hide';'+1 i:, mil) I is close to 9i・I, and the isoelectric point pi of BTG-2 is 9fi・J
It is close, and BTG3's consideration of is close to 98 phrases.
）分子【６：
ＳＤＳディスク電気泳動法より求めたところ、Ｂ −Ｔ
’ Ｇ−１の分子量は約３８，０００であり、ＢＴＧ２
の分子量は約４１，０００であり、ＢＴＧ−３の分子量
ｉ、を約４１，０００である。) Molecule [6: As determined by SDS disk electrophoresis, B-T
' The molecular weight of G-1 is approximately 38,000, and BTG2
The molecular weight of BTG-3 is about 41,000, and the molecular weight of BTG-3 is about 41,000.
、ｉ）ＭＴＧａｓｃと（７） 比較：
次にＢ Ｔ Ｇ ａ ｓ Ｏとモルモット肝由来のトラ
ンスグルタミナーゼ（ＭＴＧａｓｅ）との性質を比較Ｊ
る。尚、ＭＴＧａｓｅは、特開＠ ５８−１４９６４５
号に記載された方法で調製した。, i) Comparison of MTGasc and (7): Next, we compared the properties of BTG as O and guinea pig liver-derived transglutaminase (MTGase).
Ru. In addition, MTGase is JP-A-58-149645
It was prepared by the method described in No.
表−４には各酵素化学的性質の比較を、表−５にはＣａ
２”の活性に及ぼす影響を示す。表−４および表−５Ｊ
：り明らかのように従来主としＣ研究されでいるＭ Ｔ
Ｇ ａ ｓ ｅと放線菌由来のＦ３ＴＧａＳＣどには
酵素化学的性質において種々の差が見られ、特に温度安
定性、分子量、′Ｓ電点、↓Ｊ質特巽性に差が見られる
。また、ｃ ｉ１２４の存在］・及び：Ｊｌ存でｆ下に
４３いても本発明で使用する１３ＴＧａｓＣは作用する
点等でし明らかなＸ−がみられる。従って、本発明の各
１１まＭＴＱａｓｅと（よくの性７′■を異にするしの
ど考えられる。Table 4 shows a comparison of the chemical properties of each enzyme, and Table 5 shows Ca
Table 4 and Table 5J show the influence on the activity of 2”.
:As is clear, M T which has been studied mainly in the past
There are various differences in enzymatic chemical properties between Gase and actinobacteria-derived F3TGaSC, particularly in temperature stability, molecular weight, S electric point, and J quality characteristics. Furthermore, even if 13TGasC used in the present invention is present under f in the presence of ci124] and :Jl, a clear X- is seen in terms of its action. Therefore, it is conceivable that each of the features of the present invention may be different from MTQase.
表
表−５
（４）［３ＴＧａｓｅの？［ｉＭ
ａ） ＢＴＧ−１の製）告
ストレプトベルチシリウム・モバラエンス［＋ＴＱ １
３８１９を培地組成ポリペプトン０．２％、グリコース
Ｑ、５％、リン酸二カリウム０．２％、硫酸マグネシウ
ム０１％からなる培地（ｐＨ７１２００ｒｄ、に接種し
、３０℃、４８時間培養し、１りられた種培展液をポリ
ペプトン２．０％、シスターゲン２４０％、リン酸二カ
リウム０．２％、硫酸マグネシウム０．１％、酵母エキ
ス０．２％、消泡剤としてアデカノール（商品名、旭電
化社製品） ０．０５％からなる培地２０ｆ （Ｉ）Ｈ
７）に加え３０℃で３日間培４後ろ過し、培養液１８．
！Ｍ！ｌ！７た。このものの活性は、０．３５ｕ／ｄ
テする。Table 5 (4) [3TGase? [iM a) Produced by BTG-1) Streptoverticillium mobaraens [+TQ 1
3819 was inoculated into a medium (pH 71200rd) consisting of 0.2% polypeptone, 5% glycose Q, 0.2% dipotassium phosphate, 01% magnesium sulfate, and cultured at 30°C for 48 hours. The seed culture solution was mixed with 2.0% polypeptone, 240% cystagen, 0.2% dipotassium phosphate, 0.1% magnesium sulfate, 0.2% yeast extract, and Adecanol (trade name, Asahi) as an antifoaming agent. Denkasha product) Medium 20f consisting of 0.05% (I)H
In addition to 7), culture solution 4 was cultured at 30°C for 3 days, filtered, and culture solution 18.
! M! l! 7. The activity of this substance is 0.35u/d
Te.
培養液を塩酸でＩ）Ｎ６．５に調整し、予め００５Ｍリ
ン酸緩衝液（ＩＩＬＩＧ、５）で平衡化しておいたＣＧ
５０（商品名、オルガノ社製品）のカラムに通した。The culture solution was adjusted to I)N6.5 with hydrochloric acid, and CG was equilibrated in advance with 005M phosphate buffer (IILIG, 5).
50 (trade name, Organo Co., Ltd. product) column.
この操作でトランスグルタミナーゼは吸着さ゛れた。Transglutaminase was adsorbed by this operation.
さらに同緩衝液で不純蛋白質を洗い流した後、さらに０
．０５〜０．５ｊｖｌの同緩衝液の濃度匂配をつくり、
通液して溶出液を分画回収し、比活性の高い分画を集め
た。電導度をｌ０ｍ５以下になるように希釈後ブルーヒ
ファロースのカラムに通した。この操作でトランスグル
タミナーゼ【よ吸着された。更に００５Ｍリン酸緩衝液
（ｐｌ−１７）で不ＭＡ蛋白質を洗い流した＋９．０〜
１Ｍの食Ｆ：Ａａ度匂配をつくり通液して溶出液を回収
し比活性の高い両分を集めた。ＵＦ６０００膜を使い濃
縮し、０．５Ｍの食塩を含む０，０５Ｍリン酸緩衝液（
ＤＩ−１７）で緩衝液を用いで平衡化さ１４７こ 。Furthermore, after washing away impure proteins with the same buffer solution,
．． 05 to 0.5 jvl of the same buffer solution,
The eluate was fractionated and collected, and fractions with high specific activity were collected. After dilution so that the conductivity was 10 m5 or less, it was passed through a blue hyphalose column. Through this operation, transglutaminase was adsorbed. Further, non-MA proteins were washed away with 005M phosphate buffer (pl-17) +9.0~
A 1M food F:Aa concentration was prepared and the solution was passed through the solution, the eluate was collected, and both fractions with high specific activity were collected. Concentrate using UF6000 membrane and prepare 0.05M phosphate buffer containing 0.5M sodium chloride (
Equilibrate with buffer at DI-17).
１５１られた（Ｑ縮液を同級ｖｆＪ液で予め平衡化して
おいたロファデツクスＧ −７５（ファルマシアファイ
ンケミカル社製）を含むフコラムに通し、同緩衝液を流
１ノで溶出液を分画した。この結果粘性画分（よ単一の
ピークとして溶出された。このものの比活性は、培養ろ
液に対し６２５倍であり、回収率は４７％であった。The 151 (Q condensate) solution was passed through a fucolumn containing Lofadex G-75 (manufactured by Pharmacia Fine Chemicals) that had been equilibrated with the same grade VFJ solution, and the eluate was fractionated by passing the same buffer solution once. As a result, the viscous fraction was eluted as a single peak. The specific activity of this fraction was 625 times that of the culture filtrate, and the recovery rate was 47%.
ｂ）１３ＴＧ−・２の製造
ＢＴＧ−１の場合と同様にして、ストレプトベルブ−シ
リウム・グリロオカルネウムＩＦＣＭ２７７Ｇを３０℃
で３日間培養侵ろ過し、培養液１９ｊ！を）！Ｉた。b) Production of 13TG-・2 In the same manner as in the case of BTG-1, Streptoberbucilium grillocarneum IFCM277G was heated at 30°C.
After culturing and filtering for 3 days, the culture solution was 19j! of)! I was.
このものの活性は０．２１／戒であった。The activity of this product was 0.21/Kai.
１３７Ｇ−１の場合と同様な方法でＭ索を精製して、Ｓ
ＤＳディスク電気泳動で弔−の酵素をえた。The M chord was purified in the same manner as in the case of 137G-1, and S
The enzyme was obtained by DS disk electrophoresis.
Ｃ）Ｉ−３ＴＧ−３の製造
ＢＴＧ−１の場合と同様にして、ストレフトヘルｆシリ
ウム・シナモネウム・サブ、エスピー。C) Production of I-3TG-3 In the same manner as in the case of BTG-1, Strefther f Cirium cinnamoneum sub, sp.
シナモネウムＩ Ｆ ０１２８５２を３０℃で３日培俺
侵ろ過し、培養液１８．５でを得た。このものの酵素活
性は０５ｕ／−であった。Cinnamonium I F 012852 was cultured at 30°C for 3 days and filtered to obtain a culture solution of 18.5%. The enzyme activity of this product was 05u/-.
ＢＴＧｌの場合と同様な方法′ｃＭ木を精製して、ＳＯ
Ｓディスク電気泳動で甲−のｌ！ｙ累を得た。A method similar to the case of BTGl'cM tree is purified and SO
S-disk electrophoresis shows A-l! I got y cumulative.
以）Ｓ、本発明を実施例により更に説明する。Hereinafter, the present invention will be further explained with reference to Examples.
実施例１
九人Ｘ１ｌｏ／（ｇを水に一晩浸）へした後、２０１の
水を加えながら磨砕機ですって「ご」を］９だ。これに
消泡剤（脂肪酸モノグリセリド）を２００ｇ加えて、全
体Ｓ４Ｊとなるように加水した１１２煮釜中で加熱した
。加熱は５９閤かけて １００℃となるようにし、その
侵３分聞そのままの温度で保った。加熱終ｒ後１遇して
豆乳的４５１を得た。Example 1 Nine people x 1 lo/(soaked 9 g in water overnight), then grinded with a grinder while adding 201 g of water to 9 g. 200 g of an antifoaming agent (fatty acid monoglyceride) was added to this, and the mixture was heated in a 112 boiler to which water had been added so that the total amount was S4J. It was heated to 100°C over 59 minutes, and the temperature was maintained for 3 minutes. After heating was completed, soybean milk 451 was obtained.
豆乳は室温放置により６５℃まで冷却し、そこへグルコ
ノデルタラクトンを豆乳１１あたり３ｔＪ、ＢＴＧ−１
（比活性２．０ユニツト／＋１１９＞を豆乳１１あたり
３５０■添加した復成形器（１４ｃｍ Ｘ １６ｃｍ
ＸＡｃｍ）に充てノｖして１時間ｔＩ装置した。対照
とする系にはｒめ加熱失活しであるＢ Ｔ Ｇ −、−
１を用いた。The soy milk was cooled to 65°C by leaving it at room temperature, and glucono delta lactone was added to it at 3 tJ per 11 soy milk, BTG-1.
(Specific activity 2.0 units/+119) was added in a reshaping machine (14cm x 16cm
XAcm) was added to the tI apparatus for 1 hour. The control system contains B T G −, −, which is inactivated by heating.
1 was used.
各々の豆腐を品温が９０℃となるように加熱し、そのま
ま１０分間保った。加熱終了後、成形器ごと水中にとり
冷却し、成形器より豆腐を取り出し、流水中に３時間さ
らしてから１辺の艮ざが１．５ｃｚの立方体状にカット
し、豆腐５０９と水１００ｄをしＩ〜ルト用パウヂに充
てんした。Each piece of tofu was heated to a temperature of 90°C and kept as such for 10 minutes. After heating, the tofu was placed in water with the molding machine to cool it, and the tofu was taken out from the molding machine and exposed to running water for 3 hours, and then cut into cubes with a side length of 1.5 cz. It was filled into a pouch for I~ route.
以上のようにして調製した試料に対して日阪製作所曲ｙ
Ｊｉ！’５温高圧調理殺菌装置を用いて１１０℃でＦｏ
ＩＥが４．０となるまで処理した。又、市販絹ごし豆腐
（（０１ｉＩ橋商店製）に対しても、１記と同様にカッ
ト、充てん、レトルト処理を加えたものを調製した。For the sample prepared as above, Hisaka Seisakusho y
Ji! Fo
Processing was performed until IE reached 4.0. In addition, commercially available silken tofu (manufactured by 01iI Hashi Shoten) was cut, filled, and retorted in the same manner as in 1.
以上各々のレトルト処理済豆腐に対して、前述の市販絹
ごし豆腐（リトル１−処■！Ｉ！ｉ”＋η）を」シトロ
ール（評点Ｏ）として官能評（ｉｔｔｉ（デクスチＶ−
プロファイル法、パネル１５名）を行い表−６に示す結
果を１！′７ｋ。For each of the above-mentioned retort-processed tofu, the above-mentioned commercially available silken tofu (Little 1-doko ■!I!
The profile method (15 panelists) was performed and the results shown in Table 6 were 1! '7k.
この結果Ｊ：す、ＢＴＧａ ｓｅを作用させて調製した
レトルト処理’Ｊ！ Ｍ豆腐は一１ントロールと同等の
なめらかさ、やわらかさ、好ましさを有しているが、Ｂ
Ｔ Ｇ ａ Ｓ Ｇを作用させずに調製した対照の豆
腐及び市販絹ごし豆腐に対してしＩ−シト処理し／、：
４：）のは、かたくてぼそぼそとしＩζ豆腐として好ま
れない食感になっていることが明らかとなった。As a result, retort treatment 'J! M tofu has the same smoothness, softness, and taste as 11 tofu, but B
Control tofu prepared without TGaSG and commercially available silken tofu were treated with I-cyto:
It became clear that the texture of 4:) was hard and crumbly, which was not preferred as Iζ tofu.
表−６し１−ルｌ〜処理済豆腐の官能評価結果３−２−
１０＋ｌト２＋３
Ｏ−ＯＢ１Ｇａ５ｅ未処理十レトル１〜処理−−・
ＢＴＧａｓｅ処理１−レトルト処理Δ−−Δ 重数絹ご
し豆腐士レトルト処理実施例２
豆乳粉末（日本タンパク工業■製「ハイプロトンＪ）ｆ
３５ｇに対して水６００７を加えて、撹拌しながらガス
レンジで加熱し、沸！！後火を一定の強さに弱めて３分
間保った後人からおろした。Table 6 Shi1-ru l ~ Sensory evaluation results of processed tofu 3-2-
10+1 2+3 O-OB1Ga5e Untreated 10+1 ~ Processed--・
BTGase treatment 1-Retort treatment Δ--Δ Multiple silk tofu retort treatment Example 2 Soy milk powder (“Hyproton J” manufactured by Nippon Protein Industry ■) f
Add 6007 water to 35g, heat in a gas range while stirring, and bring to a boil! ! After reducing the heat to a certain level and keeping it for 3 minutes, it was removed from the body.
撹拌を続けながら７０℃まで冷却し、そこヘゲルー１ノ
ｆルタラクトン２ｇ、１３ＴＧ−１（比活性２．０ユニ
ツト／Ｉｌ＋！Ｆ） ３２５＃＋９を水５０ｍに溶解
させて添加した。なお、対照とする系には予め加熱失活
しである１３ＴＧ−１を用いた。The mixture was cooled to 70° C. while stirring, and 2 g of Hegel-1N rutalactone and 13TG-1 (specific activity 2.0 units/Il+!F) 325#+9 were dissolved in 50 ml of water and added thereto. Note that 13TG-1, which had been heat-inactivated in advance, was used as a control system.
上記の豆乳液を直ちにケーシングチューブ（呉羽化学工
業■製、おり巾４７＃）に充てんし、５５℃の水浴中で
３０分間又は至温で１時間放置した後、９０℃の水浴中
で３０分間加熱した。これを流水中にとり冷却した後、
日限製作所■製高渇高圧調理殺菌装置を用いて１１０℃
でＦｏ値４．０までレトルト処理した。Immediately fill a casing tube (manufactured by Kureha Kagaku Kogyo ■, width 47#) with the above soy milk solution, and leave it in a 55℃ water bath for 30 minutes or at the lowest temperature for 1 hour, and then in a 90℃ water bath for 30 minutes. Heated. After cooling it in running water,
110℃ using a high-drying high-pressure cooking sterilizer manufactured by Nichimitsu Seisakusho ■
The sample was retorted to a Fo value of 4.0.
レトルト処理後は室温まで冷ＩＩ Ｉ、、試料を３ＣＩ
Ｒずつ切り、レオメータ−での物性測定に供した。After retorting, cool the sample to room temperature.
The sample was cut into R sections and subjected to physical property measurements using a rheometer.
レオメータ−の測定条件としては、φ ７＃の球形ブラ
ンシト−を用い、試料台の上背速度５ｃｍ１分として行
い、試料豆腐の破断強度（！ｌ？／Ｊ）、変形率（％）
を求めた。なＪ３市販品絹ごし豆腐（実施＠１の中に出
てくるものと同じ）についても前述の物Ｍｉｌｌ定用試
斜と同じ形状に切り扱き上記の物性測定に供した。The rheometer measurement conditions were as follows: A 7# spherical blank was used, the upper back speed of the sample stage was set at 5 cm for 1 minute, and the breaking strength (!l?/J) and deformation rate (%) of the sample tofu
I asked for J3 commercially available silken tofu (same as that mentioned in Example 1) was also cut into the same shape as the aforementioned Mill standard test slope and subjected to the above physical property measurements.
以上の結果を表−７に示した。この結果より、ＢＴＧａ
ＳＯを作用ざゼたレトルト処理済豆腐の物性は、レトル
ト処理をしていない市販絹ごし豆腐とほぼ同等で必るが
、ｔ３Ｔ Ｇ ａ ｓ ｅを作用させていないレトルト
処理済豆腐では破断強度が上背した、寸なわら固くなっ
ていることが明らかとなった。又、両者の食感を比べる
と、ＢＴＧａｓｅを作用させた方はやわらかくなめらか
て・あるのに対し、作用させていない方は固くてぼそぼ
そとしていた。The above results are shown in Table-7. From this result, BTGa
The physical properties of retorted tofu treated with SO are almost the same as commercially available silken tofu that has not been treated with SO, but the breaking strength of retorted tofu that has not been treated with t3T Gas is higher. He turned his back, and it became clear that he had become a little stiff. Also, when comparing the textures of the two, the one treated with BTGase was soft and smooth, while the one not treated with BTGase was hard and lumpy.
表−７レトルト処理済豆腐の物性
実施例３
分耐人’１７ ＜ｌｉ白（味の素側製［アジア１コン３
２１ ） ２００！７に水２０００ノ屁と人立浦 １
００Ｕを加え軽く分散後、Ｌ３ Ｔ Ｇ −１０，０２
７硫酸カルシウム（２水塩） １４．５９を添加してか
らリイレントカツター′以下鎮臼）
ついで９０℃で１０分間加熱し＼１腐を１９だ。Table 7 Physical properties of retort processed tofu Example 3 Minute resistance '17 <li white (manufactured by Ajinomoto [Asia 1 Con 3
21) 200!7 water 2000 farts and Hitachiura 1
After adding 00U and lightly dispersing, L3 T G -10,02
7 Calcium sulfate (dihydrate salt) 14.59 was added, then the reillent cutter was crushed) Then heated at 90℃ for 10 minutes to reduce the rot to 19.
この豆腐を成形器より取り出し、水さらししてから、約
３ｃｍ角に切断した豆腐とこの豆腐に対して２倍猶の水
をレトルト用パウブに充１また。これを日阪製作所碍木
製高潟高圧調理殺菌賃首を用いＵ １２１°ＣでＦｏ
値が６０となるまで加ｊ玉１１８１’ｌ！を行った。Remove this tofu from the molding machine, soak it in water, then cut the tofu into approximately 3cm squares and fill a retort pouch with twice as much water as the tofu. This was heated to 121°C using a high-pressure cooking sterilizer made by Hisaka Manufacturing Co., Ltd.
Add 1181 balls until the value reaches 60! I did it.
冷却（ｐ得られた高渇高１処理済みの＼）腐は、はとん
どし［−シト萌の物性を保持した。The cooled (high-drying, high-1-treated \) rot obtained retained the physical properties of Hatondoshi [-Sitomoe].
実施例４
実施例１でＪ興した、Ｌ３ＴＧａｓｅを作用さゼたレト
ル１へ処理層豆腐をパウブに入ったまま２５℃、相対Ｉ
′！ｌａ度６０％の恒温器に保蔵し、１ケ月後、３ケ月
後、６ケ月後に取り出して、実施例１と同じ官能評価を
行った。その結果を表−８に示した。Example 4 The treated tofu was placed in the retort 1 treated with L3TGase in Example 1 at 25°C, relative I
′! The samples were stored in a thermostat at 60% LA, taken out after 1 month, 3 months, and 6 months, and subjected to the same sensory evaluation as in Example 1. The results are shown in Table-8.
上記と同様のことを市販の絹ごし豆腐及び用冷蔵の長期
保存豆腐（斬潟乳工業製、細雪）に対して行ったところ
、両茜とも１り列以内に腐敗をとしなった著しい変化を
おこし、喫食不能な状態となった。When the same process as above was performed on commercially available silken tofu and long-term refrigerated tofu (manufactured by Zangata Dairyu Kogyo, manufactured by Saiyuki), significant changes such as rot within one row occurred in both cases. , and became inedible.
この結Ｗ Ｊ：す、本発明の方法を用いると、調製直後
から６ケ月後までは安定した品質を保持することができ
た。By using the method of the present invention, stable quality could be maintained from immediately after preparation until 6 months later.
表−８良明保存後の官能ｉ、’Ｐ４＋咀９．宋−３−２
−１０←１ −２ １３
総合的食感
Δ□Δ
口□ａ
１ケ月後
３ケ月後
６ケ月後Table-8 Functional i after storage in Yoshiaki, 'P4 + Chew 9. Song Dynasty-3-2
-10←1 -2 13 Overall texture Δ□Δ Mouth □a After 1 month After 3 months After 6 months
Claims (1)
でペプチド鎖内のグルタミン残基のγ−カルボキシアミ
ド基のアシル転移反応を触媒する新規トランスグルタミ
ナーゼとを作用させて豆腐を調製し、このようにして調
製した豆腐を耐熱容器に充填し、レトルト処理すること
を特徴とする長期常温保存可能な豆腐の製造法。Tofu is made by reacting soymilk with a coagulant and a novel transglutaminase that catalyzes the acyl transfer reaction of the γ-carboxyamide group of the glutamine residue in the peptide chain in a Ca^2^+-independent manner at 80°C or below. A method for producing tofu that can be stored at room temperature for a long period of time, which comprises filling the thus prepared tofu into a heat-resistant container and subjecting it to retort treatment.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63219703A JP2536086B2 (en) | 1988-09-02 | 1988-09-02 | Manufacturing method of tofu that can be stored at room temperature for a long time |
| US07/401,831 US5055310A (en) | 1988-09-02 | 1989-09-01 | Process of preparing shelf-stable "tofu" at normal temperature for long term |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63219703A JP2536086B2 (en) | 1988-09-02 | 1988-09-02 | Manufacturing method of tofu that can be stored at room temperature for a long time |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH0269155A true JPH0269155A (en) | 1990-03-08 |
| JP2536086B2 JP2536086B2 (en) | 1996-09-18 |
Family
ID=16739644
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP63219703A Expired - Fee Related JP2536086B2 (en) | 1988-09-02 | 1988-09-02 | Manufacturing method of tofu that can be stored at room temperature for a long time |
Country Status (2)
| Country | Link |
|---|---|
| US (1) | US5055310A (en) |
| JP (1) | JP2536086B2 (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0631733A1 (en) | 1993-07-02 | 1995-01-04 | Ajinomoto Co., Inc. | Enzymic agent for improving the tenderization of meat |
| WO1995023524A1 (en) * | 1994-03-04 | 1995-09-08 | Tetra Lavel Holding & Finance S.A. | Process for producing protein gel |
| JPH11221039A (en) * | 1997-12-03 | 1999-08-17 | Kikkoman Corp | Production of filled bean curd |
| CN1066925C (en) * | 1994-03-04 | 2001-06-13 | 泰特拉·雷沃金融控股公司 | Process for producing protein gel |
| KR100578319B1 (en) * | 2003-12-26 | 2006-05-11 | 후지 세유 가부시키가이샤 | Soybean milk squeezed at raw materials and products thereof |
| US8176804B2 (en) | 2005-01-14 | 2012-05-15 | Nidec Sankyo Corporation | Motor actuator and opening/closing device |
| JP2021016346A (en) * | 2019-07-19 | 2021-02-15 | 大豆エナジー株式会社 | Soybean curd, raw soybean curd, food, and method for suppressing pore occurrence of soybean curd |
Families Citing this family (31)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2849773B2 (en) * | 1990-08-27 | 1999-01-27 | 天野製薬株式会社 | Method for producing transglutaminase from Streptomyces |
| CA2094570A1 (en) * | 1992-05-27 | 1993-11-28 | Jerome F. Trumbetas | Enzymatic protein process and product |
| DE69326101T2 (en) * | 1992-06-02 | 2000-04-20 | Ajinomoto Kk | Process for the production of a shaped bound food |
| JP3651003B2 (en) * | 1994-10-11 | 2005-05-25 | 味の素株式会社 | Stabilized transglutaminase and enzyme preparation containing the same |
| JP3387267B2 (en) * | 1994-10-26 | 2003-03-17 | 味の素株式会社 | Method for producing cheese using transglutaminase |
| JP3545148B2 (en) * | 1996-01-08 | 2004-07-21 | 味の素株式会社 | Edible microcapsules and foods containing the same |
| JP3637718B2 (en) * | 1996-04-10 | 2005-04-13 | 味の素株式会社 | Chocolate production method |
| JP3407599B2 (en) * | 1996-07-01 | 2003-05-19 | 味の素株式会社 | Enzyme preparation for adhesion and method for producing adhesive food |
| JP3635801B2 (en) * | 1996-08-01 | 2005-04-06 | 味の素株式会社 | Milk whey protein-containing powder and processed food using the same |
| JP3582265B2 (en) * | 1996-11-28 | 2004-10-27 | 味の素株式会社 | Modified flour and processed flour using the same |
| US6042851A (en) * | 1997-12-03 | 2000-03-28 | Kikkoman Corporation | Process for producing packed tofu |
| JP3867261B2 (en) | 1998-04-08 | 2007-01-10 | 味の素株式会社 | Enzyme preparation and method for producing noodles |
| DE10141166A1 (en) * | 2001-08-22 | 2003-03-13 | Fraunhofer Ges Forschung | Process for the production of transglutaminase cross-linked proteins of vegetable origin, protein gels and their use |
| CN1303903C (en) * | 2002-02-04 | 2007-03-14 | 不二制油株式会社 | Tofu products tolerant to freezing and process for producing the same |
| JP3824082B2 (en) * | 2003-03-13 | 2006-09-20 | 孝明 武部 | Method for producing soy milk card |
| US6908634B2 (en) * | 2003-03-20 | 2005-06-21 | Solae, Llc | Transglutaminase soy fish and meat products and analogs thereof |
| EP1731041A4 (en) * | 2004-03-30 | 2009-07-22 | Fuji Oil Co Ltd | Process for producing soybean protein and process for producing processed meat food using the soybean protein |
| US20100316764A1 (en) * | 2009-06-10 | 2010-12-16 | Engrain, LLC | Flour supplement compositions and methods for preparing wheat flour |
| KR101175152B1 (en) | 2010-01-15 | 2012-08-20 | 계명대학교 산학협력단 | Manufacturing method of whole soybean curd |
| AT11716U3 (en) * | 2010-11-12 | 2011-12-15 | Feel Good Handel Gmbh | INCREASED DURABILITY OF TOFU IN FILM PACKAGING WITHOUT COOLING |
| KR20230022455A (en) | 2011-07-12 | 2023-02-15 | 임파서블 푸즈 인크. | Methods and compositions for consumables |
| US20140220217A1 (en) | 2011-07-12 | 2014-08-07 | Maraxi, Inc. | Method and compositions for consumables |
| RU2630495C2 (en) * | 2011-07-12 | 2017-09-11 | Импоссибл Фудз Инк. | Methods and compositions for articles of consumption |
| US10039306B2 (en) | 2012-03-16 | 2018-08-07 | Impossible Foods Inc. | Methods and compositions for consumables |
| RU2557295C2 (en) * | 2012-11-15 | 2015-07-20 | Виталий Александрович Юстус | Soya-based product manufacture method |
| KR20240042553A (en) | 2013-01-11 | 2024-04-02 | 임파서블 푸즈 인크. | Methods and compositions for affecting the flavor and aroma profile of consumables |
| WO2014110540A1 (en) | 2013-01-11 | 2014-07-17 | Maraxi, Inc. | Non-dairy cheese replica comprising a coacervate |
| KR101435785B1 (en) * | 2013-04-08 | 2014-08-29 | 주식회사 풀무원 | Soybean curd using high concentration soymilk with enhanced hardness and elasticity and the preparation method thereof |
| FR3019004B1 (en) * | 2014-03-28 | 2017-10-20 | Onyx Dev Sas | FOOD COMPRISING ESSENTIALLY PLANT PROTEINS AND PROCESS FOR PREPARING THE SAME |
| KR20160140790A (en) | 2014-03-31 | 2016-12-07 | 임파서블 푸즈 인크. | Ground meat replicas |
| AT521409B1 (en) * | 2018-07-12 | 2020-11-15 | Feel Good Handel Gmbh | tofu |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS59151A (en) * | 1982-06-25 | 1984-01-05 | Agency Of Ind Science & Technol | Hologram recording material |
| JPH0789865B2 (en) * | 1986-04-28 | 1995-10-04 | 藤沢薬品工業株式会社 | Novel yogurt-like food coagulant and novel yogurt-like food using the same |
-
1988
- 1988-09-02 JP JP63219703A patent/JP2536086B2/en not_active Expired - Fee Related
-
1989
- 1989-09-01 US US07/401,831 patent/US5055310A/en not_active Expired - Lifetime
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0631733A1 (en) | 1993-07-02 | 1995-01-04 | Ajinomoto Co., Inc. | Enzymic agent for improving the tenderization of meat |
| WO1995023524A1 (en) * | 1994-03-04 | 1995-09-08 | Tetra Lavel Holding & Finance S.A. | Process for producing protein gel |
| CN1066925C (en) * | 1994-03-04 | 2001-06-13 | 泰特拉·雷沃金融控股公司 | Process for producing protein gel |
| JPH11221039A (en) * | 1997-12-03 | 1999-08-17 | Kikkoman Corp | Production of filled bean curd |
| KR100578319B1 (en) * | 2003-12-26 | 2006-05-11 | 후지 세유 가부시키가이샤 | Soybean milk squeezed at raw materials and products thereof |
| US8176804B2 (en) | 2005-01-14 | 2012-05-15 | Nidec Sankyo Corporation | Motor actuator and opening/closing device |
| JP2021016346A (en) * | 2019-07-19 | 2021-02-15 | 大豆エナジー株式会社 | Soybean curd, raw soybean curd, food, and method for suppressing pore occurrence of soybean curd |
Also Published As
| Publication number | Publication date |
|---|---|
| JP2536086B2 (en) | 1996-09-18 |
| US5055310A (en) | 1991-10-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JPH0269155A (en) | Production of soybean curd preservable for a long period at normal temperature | |
| US5156956A (en) | Transgultaminase | |
| JP5616216B2 (en) | Method for protein modification by enzyme | |
| HUE033091T2 (en) | Dairy product and process for production thereof | |
| JPH08173032A (en) | Production of cheese using transglutaminase | |
| CN107734973A (en) | Method | |
| JP3265672B2 (en) | Method for producing freeze-resistant tofu | |
| Khalid et al. | Partial purification and characterization of an aminopeptidase from Lactobacillus helveticus CNRZ 32 | |
| JP2572716B2 (en) | Novel transglutaminase | |
| El Soda et al. | Lipolytic activity of cheese related microorganisms and its impact on cheese flavour | |
| JP2611408B2 (en) | General-purpose material for paste-like food and method for producing paste-like food | |
| JP2594340B2 (en) | How to make cheese food | |
| JP2000000088A (en) | Glutaminase activity-enhanced microorganism-cultured material and its use | |
| JP2540919B2 (en) | Fried fried manufacturing method | |
| JP2556109B2 (en) | Material for meat grain | |
| JP2801376B2 (en) | Method for producing cheese flavor composition | |
| JPH08224063A (en) | Gelatinized composition of protein | |
| JP2650366B2 (en) | Solid fat and its production method | |
| JP2808752B2 (en) | Manufacturing method of retort mapo tofu that can be stored for a long time at room temperature | |
| JPH02257831A (en) | Vegetable protein powder and production of bean curd using the same | |
| US3482997A (en) | Making cheese using a modified bacterial enzyme complex from the genus bacillus | |
| JP2590373B2 (en) | New surimi and its manufacturing method | |
| JPH0343080A (en) | Production of novel transglutaminase derived from streptoverticillium | |
| JPH09262064A (en) | Soybean processed food | |
| JP2533365B2 (en) | Method of manufacturing fish surimi |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| LAPS | Cancellation because of no payment of annual fees |